Anti f4 80

pCMV-luc-C/EBPδ3′UTR were purchased from GeneCopoeia. A plasmid containing human C/EBPδ cDNA and full-length 3′UTR was kindly provided by Dr Esta Sterneck (NCI/NIH). Using this plasmid as a template, PCR was performed to amplify the fragments of the human C/EBPδ 3′UTR, and these were inserted into pCMV-luc-control vector by EcoR I/Xho I. MCPIP1 rabbit polyclonal antibody was from GeneTex (GTX110807). C/EBPβ, C/EBPδ, C/EBPα, MALT1, BCL10, A20, CYLD, RelB and β-actin antibodies were purchased from Cell Signaling Technology. LPS (L3129, O127:B8) was purchased from Sigma. SiRNAs targeting MCPIP1 and MALT1 were from Santa Cruz Biotechnology. qPCR primers were ordered from IDT. MI-2, anti-F4/80 and anti-Gr1 antibodies were purchased from R&D Systems.

Immediately after mice were sacrificed, tumor tissue was fixed in 10% formalin before paraffin embedding. Standard procedures were used for hematoxylin and eosin (H&E) staining. For immunostaining, formalin-fixed, paraffin-embedded tissue was treated with xylene, rehydrated with ethanol, and heated in a microwave with citric buffer to retrieve antigens. For blocking purpose, the tissues were incubated for 30 minutes, with 5% bovine serum albumin buffer. Followed by overnight incubation at 4° C, with primary antibodies: anti-CD3 antibody, anti-CD4 antibody, and anti-NKp46 antibody (Abcam, Cambridge, United Kingdom) at 1:100 dilutions, anti-CD8 antibody (Novus Biologicals, Minneapolis, MN) at 1:20 dilution, anti-CD31 (Novus Biologicals, Minneapolis, MN) at 1:100, anti-F4/80 (R&D system Minneapolis, MN) at 1:100, anti-CD206 (R&D system Minneapolis, MN) at 1:100, as well as anti-CD80 (Novus Biologicals, Minneapolis, MN) at 1:100. After washing, tissues were incubated with fluorescence-conjugated secondary antibodies at room temperature for 1 hr. Slides were prepared with antifade mountant with 4’,6-diamidino-2-phenylindole (DAPI).